Purpose: Retinal ischemia (RI) and progressive neuronal death are sight-threatening conditions. Mitochondrial (mt) dysfunction and fusion/fission processes have been suggested to play a role in the pathophysiology of RI. This study focuses on changes in the mt parameters of the neuroretina, retinal pigment epithelium (RPE) and choroid in a porcine high intraocular pressure (IOP)-induced RI minipig model. Methods: In one eye, an acute IOP elevation was induced in minipigs and compared to the other control eye. Activity and amount of respiratory chain complexes (RCC) were analyzed by spectrophotometry and Western blot, respectively. The coenzyme Q10 (CoQ10) content was measured using HPLC, and the ultrastructure of the mt was studied via transmission electron microscopy. The expression of selected mt-pathway genes was determined by RT-PCR. Results: At a functional level, increased RCC I activity and decreased total CoQ10 content were found in RPE cells. At a protein level, CORE2, a subunit of RCC III, and DRP1, was significantly decreased in the neuroretina. Drp1 and Opa1, protein-encoding genes responsible for mt quality control, were decreased in most of the samples from the RPE and neuroretina. Conclusions: The eyes of the minipig can be considered a potential RI model to study mt dysfunction in this disease. Strategies targeting mt protection may provide a promising way to delay the acute damage and onset of RI.

• Klíčová slova
• coenzyme Q10, minipig, mitochondrial dysfunction, retinal ischemia,
• MeSH
• glaukom * metabolismus   MeSH
• ischemie metabolismus   MeSH
• karcinom z renálních buněk * metabolismus   MeSH
• miniaturní prasata MeSH
• mitochondrie metabolismus   MeSH
• nádory ledvin * metabolismus   MeSH
• nitrooční tlak MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The Acyl-CoA-binding domain-containing protein (ACBD3) plays multiple roles across the cell. Although generally associated with the Golgi apparatus, it operates also in mitochondria. In steroidogenic cells, ACBD3 is an important part of a multiprotein complex transporting cholesterol into mitochondria. Balance in mitochondrial cholesterol is essential for proper mitochondrial protein biosynthesis, among others. We generated ACBD3 knock-out (ACBD3-KO) HEK293 and HeLa cells and characterized the impact of protein absence on mitochondria, Golgi, and lipid profile. In ACBD3-KO cells, cholesterol level and mitochondrial structure and functions are not altered, demonstrating that an alternative pathway of cholesterol transport into mitochondria exists. However, ACBD3-KO cells exhibit enlarged Golgi area with absence of stacks and ribbon-like formation, confirming the importance of ACBD3 in Golgi stacking. The glycosylation of the LAMP2 glycoprotein was not affected by the altered Golgi structure. Moreover, decreased sphingomyelins together with normal ceramides and sphingomyelin synthase activity reveal the importance of ACBD3 in ceramide transport from ER to Golgi.

• Klíčová slova
• ACBD3, Golgi, OXPHOS, cholesterol, knock-out, mitochondria,
• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• biologický transport fyziologie   MeSH
• ceramidy metabolismus   MeSH
• cholesterol metabolismus   MeSH
• glykosylace MeSH
• Golgiho aparát metabolismus   MeSH
• HEK293 buňky MeSH
• HeLa buňky MeSH
• lidé MeSH
• membránové proteiny metabolismus   MeSH
• membránový protein 2 asociovaný s lyzozomy metabolismus   MeSH
• mitochondrie metabolismus   MeSH
• signální transdukce fyziologie   MeSH
• transferasy pro jiné substituované fosfátové skupiny metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ACBD3 protein, human MeSH Prohlížeč
• adaptorové proteiny signální transdukční MeSH
• ceramidy MeSH
• cholesterol MeSH
• membránové proteiny MeSH
• membránový protein 2 asociovaný s lyzozomy MeSH
• phosphatidylcholine-ceramide phosphocholine transferase MeSH Prohlížeč
• transferasy pro jiné substituované fosfátové skupiny MeSH